1. Field of the Invention
The present invention related generally to an infrared (IR) spectroscopy method for the diagnosis of a disease state of a mammal such as a human being. More particularly, the method of the invention allows for specific and quick diagnosis of a variety of pathological conditions by analyzing the IR spectra of the patient""s blood.
2. Description of the Prior Art
Diagnosis of a disease state of a human being is done using a variety of laboratory techniques. That, in conjunction with the general medical assessment of the patient""s condition and symptoms allows a modern medical practitioner to determine in most cases the accurate diagnosis of a patient. It is well known that the ease of treatment depends greatly on the speed of such diagnosis. Thus, the sensitivity and precise discriminatory nature of the early diagnostic methods is very important in the effective treatment of patients.
There is a variety of sources of biological information about the general medical state of a patient that is used in the laboratory analysis as various tissue and cell samples as well as biological fluids that may be taken from a human being. An example of a biological fluid is blood, saliva, sweat, urine, semen, various gland secretions, joint lubricant fluids, lymphatic fluids, and so on. Typically, these fluid or tissue samples come from a specific organ so that a further laboratory analysis can reveal a certain pathologic condition of that particular organ. Blood, however, represents a universal fluid circulating throughout the human body and as such provides an almost unique cumulative source of biological information about the body as a whole as well as about individual organs.
A number of laboratory techniques have been developed in the recent years to study blood and other biological fluids and tissues. Many of them are aimed specifically to identify a certain pathological condition. However, no universal technique exists today that would allow for a quick, minimally invasive broad diagnosis of various organ specific conditions.
Usually, a simple elevation of a certain particle count in blood (such as leukocytes) is indicative of some inflammation process. Also, in recent years physicians have used such serum enzymes as creatine phosphokinase, lactic dehydrogenase, and aminotransferases, which are released in large quantities into the blood from necrotic tissue, for diagnostic purposes (for example for diagnosis of myocardial infarction or hepatitis). However, elevation of the serum enzymes level, as well as an increased level of leukocytes, is not organospecific and can be used only as an additional method of diagnostics. In that case, it is up to a physician to determine the exact organ or a group of organs responsible for a pathologic condition based on the symptom analysis, which may be not present, misleading, or confusing at times.
Among methods other than IR spectroscopy the method of microparticle enzyme immunoassay technology is the closest one to the suggested by us method by its aim and problem solving. This technique is used primarily for diagnostics of acute myocardial infarction and based on determination of a cardiac form of Troponin-1, which is the regulatory submit of the troponin complex associated with the actin thin filament within muscle cells. This method also has numerous limitations because any conditions resulting in myocardial cell damage can potentially increase cardiac Troponin-1 levels. These conditions include, but are not limited to: angina, unstable angina, congestive heart failure, myocarditis, cardiac surgery or invasive testing. Thus, this method provides an opportunity to diagnose the pathology of the specific organ, in this particular case the heart, but does not allow for making the specific diagnosis, that is for determining the particular heart disease. This leads to an incorrect diagnosis or a complete misdiagnosis of a patient with obvious consequences.
IR spectroscopy is a very sensitive chemical analysis method, which is routinely used by organic chemists and biochemists as a molecular probe. When infrared light is passed through a sample of an organic compound, some of the frequencies are absorbed and some are transmitted through the sample without being absorbed. As a result, this selective light absorbance is recorded as a chart by the machine and can be further used to determine the exact chemical composition of the sample under investigation. The term xe2x80x9cIR spectroscopyxe2x80x9d is used here to include laser-Raman spectroscopy, Raman con-focal laser spectroscopy, Fourier Transform infrared spectroscopy, or any other infrared spectroscopy technique. Organic applications of IR spectroscopy are almost entirely concerned with frequencies in the range of 650-4000 cmxe2x88x921. Frequencies lower than 650 cmxe2x88x921 are called far infrared and those greater than 4000 cmxe2x88x921 are called near infrared.
There are several important advantages in using this technique: results are obtained relatively quickly with less labor input than many other diagnostic techniques; the use of IR spectroscopy may provide a more precise information on the exact nature of a disease based on sampling of blood or other biological fluid; the method also allows to monitor the dynamics of the characteristic change, which is important in determining the exact stage of the disease.
It has been established in the prior art that certain diseases such as cancer can substantially change the IR characteristic of blood and some other biological fluids. Therefore, IR can be used to determine the presence or absence of cancer cells and even to determine whether the tumor is malignant or not.
Several US and international patents contain a description of utilizing IR spectroscopy for cancer diagnosis. U.S. Pat. No. 5,261,410 by Alfano discloses a method of determining the cancerous state of a human tissue, particularly in breast tissue after exposing a sample of that tissue to IR light. This method has limitations due to the need for tissue biopsy and also since it does not allow for diagnosis of other organs.
U.S. Pat. No. 5,186,162 by Oong illustrates a method of using the IR spectroscopy of a tissue sample or a culture for differentiation between the presence of normal and cancerous cells. The specimen under investigation may be a Papanicolu smear, a cervical specimen, an endocervical specimen, or a vaginal or uterus specimen. This method allows only for cancer diagnosis and is limited to the particular organ under investigation, which has to be suspected to contain cancerous cells even before the test is done. Thus, early diagnosis, especially other than cancer, is not feasible with this technique.
PCT Patent Application No. WO 97/14961 by Antipov describes the use of IR spectroscopy means for cancer diagnosis of various organs by analyzing the IR spectra of blood. This method allows for diagnosis of malignant disorders using the method of multiple irregular total internal reflection in the IR range. It utilizes blood as a source of biological information and allows for limited determination of a specific organ that contains cancerous cells. At the same time, in addition to requiring some special IR tools and exotic evaluation technique (which are not routinely available or known in the medical laboratory community), this method is limited to the diagnosis of other cancer only and does not allow for diagnosis of other important diseases and conditions such as inflammation of a certain internal organ.
A multivariate classification techniques are applied to spectra from cell and tissue samples irradiated with IR light according to the U.S. Pat. No. 5,596,992 by Haaland in order to determine if the samples are normal or cancerous. Classification can be made using infrared spectroscopy and analysis tools such as partial least square technique (PLS), principal component regression (PCR), and linear discriminant analysis. These classifications can be used to distinguish normal, hyperplastic, and neoplastic cells. Lymphatic fluid and tissue samples are used as the object of evaluation. The important limitation of this invention is that it requires a complex apparatus and a special microscope to be used and, as described before, allows only for cancer diagnosis.
U.S. Pat. No. 5,733,739 by Zakim discloses yet another IR spectroscopy machine-based cancer diagnosis method that allows differentiating between normal and abnormal (cancerous) cells. It has all the same limitations as the patents described above such as the need for a difficult organ tissue sample collection procedure (biopsy), which can not be done routinely but rather is performed very rarely due to its traumatic nature. In addition, this method can not include other than cancer diseases of internal organs.
Other than cancer diagnostic methods have also been suggested in the prior art. PCT Patent Application No. WO 97/32194 by Ashdown discloses the use of IR spectroscopy of blood or its components for determination of cellular immunity. Fourier Transform infrared spectroscopy is a preferred method of analysis according to this invention. This invention can be used for the determination of cellular immunity in patients with immunodeficiency, autoimmunity, and contact with infectious diseases, allergies, hypersensitivity, and cancer. It is also claimed to be used for determination of tissue transplant compatibility. Although a useful technique, this method by itself does not allow for differentiation between various internal organs as being the source of the disease let alone diagnosing other inflammatory conditions.
Chemical composition of a biological fluid such as blood can be determined using a near IR spectroscopy method described in U.S. Pat. No. 4,975,581 by Robinson. This method can be used effectively to determine the glucose level in blood and thus is of interest to diabetic patients. However, a simple determination of glucose level cannot be used effectively in diabetic diagnosis and screening since no determination can be made as to whether the source of higher glucose levels is physiologically acceptable or pathologic. This method is also claimed as capable of determining other blood parameters such as alcohol presence, fatty acid content and others and as such can be used as a replacement method to the currently employed techniques used for such determinations. However, it cannot produce clinically important information about the presence of inflammatory processes in various internal organs and therefore cannot be used as a broad diagnostic tool. Hemoglobin and hematocrit concentration in blood can be determined by using a near IR spectroscopy technique suggested by Osten in U.S. Pat. No. 5,830,133. Although fast and reliable, this method is limited to just the claimed purpose of hemoglobin evaluation and does not allow for internal organ diagnosis.
Eysel in the U.S. Pat. No. 5,473,160 suggests another specific application of IR spectroscopy of synovial fluid for diagnosing arthritis disorders. Osteoarthritis is diagnosed by comparing the synovial fluid near IR spectra retrieved from the patient with that of either a known healthy or diseased spectra. The method described in the patent is limited to only arthritis diagnosis.
Finally, my Russian Patent # SU 1,698,775 describes a method of determining the level of septic state in children using IR spectra. The presence of a xe2x80x9cwavedxe2x80x9d-shape peak in the range of about 1095 cmxe2x88x921 to about 1105 cmxe2x88x921 of the same elevation as the liver portion of the chart is used as an indicator of a presepsis condition. Same at about 1080 cmxe2x88x921 points out to the initial stage of sepsis. A peak at about 1040 cmxe2x88x921 points to a medium level of sepsis, and finally a peak at about 1010 cmxe2x88x921 is associated with extended sepsis. This method is very useful but applicable only in small children and cannot differentiate between specific internal organs.
All IR spectroscopy techniques suggested in the prior art have similar limitations: they allow for selective diagnosis of some global disorders, mostly cancer, and sometimes capable of pointing to a certain internal organ. However, no universal technique exists for a broad diagnosis of organ specific disorders and the stage of their development in various internal organs, for example, a variety of inflammation conditions. Therefore, the need exists for such a broad diagnosis method.
Accordingly, it is an object of the present invention to overcome these and other drawbacks of the prior art by providing a novel machine-based method for a broad and comprehensive diagnosis of various pathological conditions of the human being by using IR spectroscopy of blood.
It is another object of the invention to provide a method for diagnosis for a variety of organ specific disorders using the IR spectra of patient""s blood.
It is another object of the present invention to provide a method for early stage diagnosis of such conditions in patients of any age when conventional chemical analysis techniques are not capable of determining the diagnosis and no or little clinical symptoms are present.
It is a further object of the present invention to provide a method of monitoring the progression of the disease; the progression and results of the treatment of that disease; the effectiveness of the treatment therapy including the effect of medication; and the determination of the final cure from the disease, such method based on IR spectroscopy of blood.
It is yet a further object of the present invention to provide a method of fast and comprehensive diagnosis of a variety of pathological conditions of a human being soon after the patient is first presented to the medical professional.
It is another object of the invention to provide a method for selecting especially healthy people for certain special needs such as fighter pilots, astronauts, and other professions requiring physical endurance.
The method of the invention is based on the discovery that various major organs and their disease state can affect the IR spectra of human blood in a very specific individual way. In other words, certain ranges of frequencies in the IR spectra are indicative of certain specific organs and their pathologies.
The physiological importance of blood flow is that it transports all necessary ingredients to the internal organs and then removes all the components that have to be discarded by these organs. Whole blood contains over 1000 chemical components. Although IR spectroscopy is a very sensitive analysis tool, it cannot pick up reliably the presence of components having less than about 5% of the total mass of the sample. In a dried blood sample, the distribution of mass is such that most of it is represented by hemoglobin ranging from about 64 to 87%. The next highest number is lipids occupying about 8.8% of the total mass. All other components are less than 5% and should not affect significantly the IR spectra of the whole blood. Of course, special techniques are available to separate certain components from the whole blood. Themselves may in turn, use their selective IR spectra for diagnostic purposes.
Hemoglobin has a very complex chemical protein structure capable of reacting to the presence in blood of a variety of compounds. Such compounds entering the blood stream may be the result of metabolic processes, sub-components of certain cells that have to be carried away from diseased organs, or some other products of pathological processes in the body. All of these complex compounds have the ability to attach to hemoglobin proteins or in some other way effect its chemical structure. Since most of pathological reactions are organ specific, it is suggested that the unique compounds released in the blood stream can be used as an organ specific diagnostic tool provided that the information can be properly extracted and analyzed. The similar process takes place by the interaction of the above-mentioned pathologic products with phospholipids of erythrocytes.
Thus, the basis of the suggested by us method of diagnostics is not the identification of products of pathological processes by their spectra. The basis of this method is the change of blood spectra of a healthy human being (frequency shift, changes in the intensity and shape of the absorbance band) under the influence of intermolecular forces, which are brought about by the interaction of the bonds found in the whole blood of a healthy human being with the bonds of the corresponding metabolites and/or the products of pathological processes, entering the blood, if a specific disorder is taking place in the body.
Whereat, the concentration of the above-mentioned metabolites and/or the products of the decay of the specific organs and tissues, not found in the normal blood, is, as a rule, significantly lower than the threshold concentration which is necessary to diagnose a disorder of a specific organ using routine biochemical methods. The method of IR spectroscopy suggested by us detects organ specific molecules that precede the appearance of a whole complex of organ specific antigens in the blood.
It is possible that the mentioned above decay products of the specific organs can be determined by using immunology or other spectroscopy methods, based on the diagnosis of specific antigens or substances. However, in variety of illnesses, some of which are appendicitis, cholecystitis, pancreatitis e.t.c. it is impossible to make a diagnosis using the antigens or substances because in most cases the nature of this antigens or substances is unknown.
Besides, it should be noted that in overwhelming majority of cases (if not in all of them), the specific substance characteristic to a certain disease enters the blood in concentrations that are significantly less than 5% of the sample specimen and can not be seen by themselves. We can see this substance only if it is involved in some kind of intermolecular interaction with the hemoglobin bonds or that of phospholipids, which make up the basis for the blood spectra of a healthy human being.
Apparently, the said changes can be observed in those parts of spectra where chemical bonds absorb, which are mostly typical of (mostly absorbing) the substance entering the blood. If in the particular region of spectra the vibrations of hemoglobin bonds and/or the bond of phospholipids of erythrocytes are not found or faintly pronounced, we will observe the appearance of new ranges of absorbance absent in the blood spectra of a healthy human being, as for example, in the case of kidney disorder. However if in the specific region of the spectra the vibrations of hemoglobin bonds and/or the bonds of erythrocyte phospholipids are well pronounced (peaks, bands), then the result of intermolecular interaction will depend on the chemical nature and the number of bonds of the substance entering the blood.
As a rule, when a small amount of the substance enters the blood, the intensity of the corresponding absorbance range will increase, as for example in the case of acute myocarditis. By contrast when a large amount of the substance enters the blood, the intensity of the absorbance range will reduce, as for example in case of myocardial infarction. If the band of absorbance in norm is well pronounced, as for example in the region in which nervous system diseases are diagnosed, then the appearance of the products of the organ decay with the similar chemical bonds will result in the reduction of the absorbance band. The larger the amount of the decay products that enter the blood, the more significant the reduction of the absorbance band will be. For example, the brain injury of different degree of the severity would correspond to the different degree of the intensity reduction.
If the absorbance of the bonds of pathological products entering the blood occurs beyond the region where the bonds of substances found in the normal blood absorb, namely, next to the said region, then we would observe the absorbance range shift, as it happens in case of allergy or hepatitis.
At the same time, it is possible that the complex molecule of hemoglobin can be a molecular system of correspondence of the body, on which the condition of internal organs and the small changes of their metabolism are represented. These changes can be detected by us due to conformational changes of hemoglobin structure. It is similar to such known correspondence systems as reflexogenic areas of skin or the iris of an eye.
According to the method of the invention, a sample of patient""s blood is taken and processed in a known way to allow for recording of IR spectra. One common way to prepare the blood sample is to dry the blood on glass plate and then remove the dried particles from the glass, mix with a filler and compress into a tablet or, in case of a liquid filler, place the suspension in a shape allowing the use of an IR recording apparatus. That example of a preparation technique is described in greater detail in my Russian Patent No. SU 1,698,775 and allows for easy transportation of the sample to the IR laboratory as well as for subsequent easy handling and storage. Once in the lab, the IR light is passed through the sample, the IR spectra of the patient""s blood is recorded and compared with the general norm from known healthy subjects as well as with the IR charts from known disease conditions. The presence or absence of certain graphical elements on the chart in comparison to the normal or known disease chart is used for a variety of organ specific diagnostic purposes described in greater detail below.
The importance of the method in the present invention is in utilization of a newly discovered fact that specific organs and their respective disease states have particular and distinct ranges of frequency in which the IR absorbance spectra changes (specific pictures). Therefore, once a change or deviation from an established norm is detected, the range of frequency where the change occurs points out to a specific organ in question. Most of the times, based on a particular frequency with a pronounced change, clinician would be able to diagnose a specific pathological condition for a particular organ. The regions responsible for individual organs and organ systems are described below progressing from lower frequencies towards higher frequencies (FIGS. 1 and 2).
REGION 1, with frequency ranging from about 600 cmxe2x88x921 to about 700 cmxe2x88x921 is indicative of various hypoxic conditions associated with reduction in oxygen availability in organism. Hypothetically, in this region out-of-plane deformation vibrations of NH-bonds of hemoglobin and OCN-turns of the peptide groups of hemoglobin are observed, which increase by lack of oxygen in blood as a result of activation of vibrations of the iron-containing group of hemoglobin""s hem.
REGION 2, with frequency ranging from about 700 cmxe2x88x921 to about 800 cmxe2x88x921, is indicative of liver failure. Hypothetically, in this region we observe deformation rocking vibrations of CH2-groups in lipids, when they enter the blood with concentration significantly exceeding the norm, which occurs only in case of liver failure.
REGION 3, with frequency ranging from about 800 cmxe2x88x921 to about 1000 cmxe2x88x921 is indicative of disorders of gastrointestinal tract, kidneys, endocrine glands, as well as some types of cancer. This region, depending on the structure of chemical compounds shown in the spectra, is divided into Subregion 3A with frequency ranging from about 800 cmxe2x88x921 to about 900 cmxe2x88x921 and Subregion 3B with frequency ranging from about 900 cmxe2x88x921 to about 1000 cmxe2x88x921.
Subregion 3A is indicative of the disorders of colon, kidneys and endocrine glands. Hypothetically, if a colon disorders exists, in the range of about 850 cmxe2x88x921 to about 900 cmxe2x88x921 we observe deformation vibrations of NH, CH and CHxe2x95x90CH bonds of some toxic products, developed in large intestine as a result of proteins putrefaction (indole, skatole, phenylpropeonic, phenylacetic, parahydroxylphenyllactic acids, etc.). These products are normally not found, but can enter blood in very low amounts as the result of colon disorders.
Hypothetically, in the case of kidney disorders, in the range of about 850 cmxe2x88x921 to about 860 cmxe2x88x921 we observe deformation vibrations of NH, CH and CHxe2x95x90CH bonds of the uric acid, urea and decay products of protein-lipids antigens from damaged kidneys cells. It happens, when the concentration of the specified above metabolites in the blood deviate from the conventional standard, determined by the IR spectra.
Hypothetically, when an endocrine disorders take place in the body, in the range of about 880 cmxe2x88x921 to about 900 cmxe2x88x921 we observe deformation vibrations of CH and CHxe2x95x90CH bonds of a number of hypophysiotropic and steroid hormones. The spectra of a healthy human being do not show these hormones.
Subregion 3B, is indicative of the disorders of stomach, pancreas, small intestine and some types of cancer. Hypothetically, if a patient suffers from stomach disorders, in the range of about 930 cmxe2x88x921 to 950 cmxe2x88x921 we observe deformation vibrations of NH and CH bonds of protein products, which are a result of stomach mucous membrane inflammation.
Hypothetically, in the area of about 940 cmxe2x88x921 we can observe deformation vibrations of NH and CH bonds of protein products, which form as a result of pancreas tissue inflammation.
Hypothetically, in case of duodenum, jejunum and ileum disorders, in the frequencies ranging from about 950 cmxe2x88x921 to about 1000 cmxe2x88x921 we observe deformation vibrations of CH and CHxe2x95x90CH bonds of phospholipid products, as well as NH and CH bonds of protein products formed by inflammation of these organs.
Hypothetically, if a patient suffers from certain types of cancer which correlate with immunodeficiency conditions, in the area of about 908 cmxe2x88x921 we observe deformation vibrations of CH and NH bonds of protein-lipid antigens and RNA from the cancerous tissue.
REGION 4, with frequency ranging from about 1000 cmxe2x88x921 to 1140 cmxe2x88x921, is indicative of the disorders of liver and immune system (different kinds of immunodeficiency and allergy). This region, depending on the functional characteristics, can be subdivided into Subregion 4A with frequency ranging from about 1100 cmxe2x88x921 to 1140 cmxe2x88x921 or more exactly the peak at a frequency of about 1130 cmxe2x88x921, and Subregion 4B with frequency ranging from about 1000 cmxe2x88x921 to 1100 cmxe2x88x921.
Subregion 4A, especially the peak at a frequency of about 1130 cmxe2x88x921, to be exact, is indicative of normal liver function. This subregion represents a spectra of SH-groups of hemoglobin, consisting of sulfur containing amino acids (cysteine, cystine and methionine) and responsible for respiratory function of hemoglobin. Also, sulfur-groups of hemoglobin determine its reactivity, that is its ability to react not only with oxygen but also with other compounds. In connection with this, when toxic metabolites, products of decay of organs and tissues enter the blood and/or the liver detoxification function is upset, the peak at a frequency of about 1130 cmxe2x88x921 disappears or modifies. This is a result of interaction between SH-groups of hemoglobin and the above-mentioned toxins, as well as of the formation of other compounds with a new conformation structure. We cannot also rule out that sulfa lipids of erythrocytes and possibly ether-sulfonic acids, which are associated with liver detoxification function, form the peak at about 1130 cmxe2x88x921. In view of this, the normal position of the peak in the range of about 1130 cmxe2x88x921 defines the normal liver condition, while its disappearance or changes in its conformation correlate with liver function disorders.
Subregion 4B is indicative of the disorders of the liver (hepatitis) and such immune disorders as allergy; T- and B-cells immunodeficiency; and nonspecific phagocytic immunodeficiency (sepsis: SU Patent No. 1,698,775).
Hypothetically, when hepatitis takes place in the frequency range of about 1030 cmxe2x88x921 to about 1000 cmxe2x88x921 we observe vibration modes of glycogen. In case of allergy in the range of about 1080 cmxe2x88x921 to about 1090 cmxe2x88x921 we observe the vibrations of Cxe2x80x94Oxe2x80x94P bonds of phospholipids from decayed cells, and Pxe2x80x94O bonds of ADP and AMP. If a patient suffers from T- and B-cells immunodeficiency associated with a tumor, we observe vibrations of Cxe2x80x94Oxe2x80x94P bonds of DNA and RNA at frequencies of about 1030 cm31 1 and 1060 cmxe2x88x921.
Hypothetically, if nonspecific phagocyte immunodeficiency (sepsis) exists, we observe vibrations of Cxe2x80x94Oxe2x80x94P and Pxe2x80x94OH bonds of all above-mentioned components, namely ADP, AMP, DNA, RNA, phospholipids, as well as vibrations of metal containing groups of cytochromes from decayed mitochondria of cells.
REGION 5, with frequency ranging from about 1140 cmxe2x88x921 to about 1350 cmxe2x88x921, is indicative of the disorders of the heart. Depending on the chemical compounds, revealed in this region of the spectrum, we subdivide this region into three subregions:
Subregion 5A, with frequency ranging from about 1140 cmxe2x88x921 to about 1200 cmxe2x88x921, primarily the peak at a frequency of about 1160 cmxe2x88x921, reflects the condition of a myocardial muscle in norm (FIG. 1);
Subregion 5B: the straight descendent under acute angle line in the range of about 1180 cmxe2x88x921 to about 1200 cmxe2x88x921, reflects the condition of the heart valves in norm and/or absence of heart insufficiency (FIG. 1), and
Subregion 5C: the area ranging from about 1200 cmxe2x88x921 through about 1350 cmxe2x88x921, including a big peak at a frequency of about 1250 cmxe2x88x921, reflects the cardiac rhythm in norm (FIG. 1).
Changes of IR absorbance in these areas reflect the appropriate pathologies of myocardial muscle, disorders of heart valves and/or presence of heart insufficiency of different degree, and cardiac arrhythmia.
Hypothetically, the peak at about 1160 cmxe2x88x921 (Subregion 5A) normally is the spectra of Cxe2x80x94Oxe2x80x94C and Pxe2x80x94Oxe2x80x94C bonds of phospholipids of erythrocytes. When Cxe2x80x94Oxe2x80x94C and Pxe2x80x94Oxe2x80x94C bonds of phospholipids from the damaged heart cells, especially cardiolipin (serinephosphatide) appears in the blood and in the spectra, it allows for diagnosis of myocard diseases and heart valves disorders, including heart failure.
Hypothetically, the peak at about 1250 cmxe2x88x921 (Subregion 5C), normally and primarily represents amid III of the hemoglobin peptid chain (NH turn in-plane of peptide chain of hemoglobin), and to the lower extend, Pxe2x95x90O and Cxe2x80x94Oxe2x80x94C bonds of phospholipids of erythrocytes. The appearance in the blood and in the spectra of Pxe2x95x90O, Pxe2x80x94OH and Cxe2x80x94Oxe2x80x94C bonds of phospholipids, and, probably, NH bonds of protein from the damaged myocard cells, in the range of about 1200 cmxe2x88x921 to about 1350 cmxe2x88x921 strictly correlates with the heart rhythm disorder, that is evident of the damage of the heart conductive system, and allows for diagnosis of various types of arrhythmia.
REGION 6 (MIXED), with frequency ranging from about 1100 cmxe2x88x921 to about 1200 cmxe2x88x921 includes confluence of Subregion 4A (which is indicative of the normal liver function) and Subregion 5A (which is indicative of the normal myocard function of the heart). Simultaneous increase of IR absorbance in this region, especially of the peaks at about 1130 cmxe2x88x921 and 1160 cmxe2x88x921, is indicative of the inflammation disorders of the upper respiratory tract and lungs. Hypothetically, that is because by the above-mentioned diseases one can see in this region Pxe2x95x90O and Pxe2x80x94OH vibrations of ATP; SS and SH vibrations of sulfolipids and also Cxe2x80x94Oxe2x80x94C and Pxe2x80x94Oxe2x80x94C vibrations of phospholipids, which appear in the blood as a result of lung cells damage and throw of ATP into the blood, which is necessary to provide energy for a general inflammation process.
REGION 7, with frequency ranging from about 1400 cmxe2x88x921 to about 1700 cmxe2x88x921, is indicative of the disorders of brain and central nervous system. Depending on the degree of severity of brain disorders, shown in the spectra, we subdivided this region into Subregion 7A, with frequency ranging from about 1400 cmxe2x88x921 to about 1500 cmxe2x88x921, or more definitely, the peaks at frequencies of about 1410 cmxe2x88x921 and 1460 cmxe2x88x921. And Subregion 7B, with frequency ranging from about 1500 cmxe2x88x921 to about 1700 cmxe2x88x921, or more definitely, the peaks at frequencies of about 1550 cmxe2x88x921 and 1650 cmxe2x88x921.
Hypothetically, the peak at a frequency of about 1410 cmxe2x88x921 (Subregion 7A) represents symmetric CO stretching vibrations and symmetric CH3 bending vibrations of hemoglobin amino acids. The peak at a frequency of about 1460 cmxe2x88x921 (Subregion 7A) is probably indicative of CH2 bending vibrations and asymmetric CH3 bending vibrations of all components of the whole dry blood.
If a brain damage take place, the appearance in the blood of the protein products of the nerve tissue decay, namely, of neuroglobulin and neurostromin, containing CO bonds of amino acids, results in the changes of the peak conformation at a frequency of about 1410 cmxe2x88x921. And the appearance of even small amounts of products of nerve cell membranes decay, containing CH2 and CH3 bonds, results in the changes of peak conformation at a frequency of about 1460 cmxe2x88x921.
On the whole, the changes in spectra in the Subregion 7A are indicative of less severe, typical for early stage of central nervous system diseases and/or functional brain disorders, such as a minor degree of intracranial hypertension, neurosis or early stage of convulsion syndrome.
Hypothetically, in Subregion 7B the peak at a frequency of about 1550 cmxe2x88x921 represents primarily asymmetric CO stretching vibrations and, secondly, NH bending and CN bending vibrations of hemoglobin amino acids. The peak at a frequency of about 1650 cmxe2x88x921 represents primarily CO bonds of a-spiral of the hemoglobin peptide chain and, secondly, CO bonds of acetalphosphatides (plasmalogens) of erythrocyte lipids and Nxe2x89xa1CH3)3 bonds of cholinephosphatides (lecithins), which are part of erythrocyte lipids.
When brain is severely damaged, the blood shows not only the products of protein decay of brain tissues, containing asymmetric CO bonds with vibrations at a frequency of about 1590 cmxe2x88x921 and NH bending vibrations at a frequency of about 1570 cmxe2x88x921. Also one can see the products of a more severe brain damages, such as a sphingomyelin with asymmetric CO and CN bonds vibrations in the frequency range of about 1550 cmxe2x88x921-1655 cmxe2x88x921. These products consist of sphingomyelin, phosphatidylserine, phosphatidylcholine and other phosphoric acids with CC-cis bonds, vibration of which one can see at frequencies of about 1660 cmxe2x88x921 and 1670 cmxe2x88x921. Since phosphatidylcholine also contain with Nxe2x89xa1(CH3)3 bonds, one can identify vibrations at a frequency of about 1630 cmxe2x88x921. On the whole, the changes in spectra in the Subregion 7B are indicative of more severe damages of brain and central nervous system, such as head trauma, hypoxic encephalopathy with hemorrhage, etc.
REGION 8, with frequency ranging from about 3000 cmxe2x88x921 to about 3100 cmxe2x88x921, especially the peak at a frequency of about 3060 cmxe2x88x921, is indicative of severe chronic infections, such as pulmonary tuberculosis. Hypothetically, the peak at a frequency of about 3060 cmxe2x88x921 primarily represents xe2x80x94NH3+ bond of associated molecules of secondary amids, constituting side chains and terminal groups of the hemoglobin protein molecules. Also, in the frequency range of about 3010 cmxe2x88x921 to about 3095 cmxe2x88x921 one can see xe2x95x90Cxe2x80x94H stretching vibration of erythrocyte phospholipids.
In the case of severe chronic infections, including pulmonary tuberculosis, the intoxication of the body is strongly pronounced, which results in products of protein decay entering the blood. These products contain xe2x80x94NH3+ bond of associated molecules of secondary amids, constituting side chains and terminal groups of protein molecules. Besides, we cannot rule out the appearance in blood of great amounts of decay products of the tuberculosis bacillus itself, particularly, of the tuberculostearic acid, containing xe2x95x90Cxe2x80x94H bonds, which absorb in the range of about 3010 cmxe2x88x921 to about 3095 cmxe2x88x921.
On the base of outlined above yet another application of the method is that it allows for monitoring the progression of the disease or the success of the treatment, by comparing subsequent blood samples spectra to the initial one.
Also, the use of the method is possible when selecting the healthiest men for the professions where extreme physical endurance is needed.
In addition the method of the invention allows for rapid and accurate early diagnosis, frequently pointing out to the initial stage of the disorder when no clinical manifestations are yet observed.
The method is also particularly useful when a physician cannot count on the patient""s response, as for example, in case of a patient being unconscious after an accident or with a pediatric patient.
Another advantage is that the method of the present invention allows for a rapid differentiation between a number of likely medical conditions, all of them presenting with similar clinical symptoms. This method has real advantages because it gives us a chance to simultaneously diagnose different organs pathology in contrast to some well known diagnostic methods such as ECG or EEG, which can be used for diagnostic diseases of only one organ (heart or brain).
The method can be used in insurance medicine, military and space medicine, forensic and veterinary medicine, pharmacology and parasitology. It can also be helpful for diagnostics of some oncology (retroperitoneal lymphoma) and infectious diseases (HIV, hepatitis B, etc.) on the earliest stage, when no clinical manifestations and antibodies are present. Moreover, the method can be used in different branches of biology (zoology, entomology, ichthyology, ornithology) and botany.